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Identifying transcriptional targets of vasopressin using combined ChIP‐seq and RNA‐seq analysis of mpkCCD cells (1181.6)
Author(s) -
Claxton J'Neka,
Sandoval Pablo,
Lee Jae Wook,
Hoffert Jason,
Knepper Mark
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.1181.6
Subject(s) - chromatin immunoprecipitation , vasopressin , biology , gene , chromatin , rna seq , rna , gene expression , microbiology and biotechnology , genomic dna , rna polymerase ii , transcriptome , promoter , genetics
The peptide hormone vasopressin regulates transport in the renal collecting duct in part through regulation of gene expression. To obtain a global profile of genes regulated by vasopressin, we performed ChIP‐seq analysis of RNA polymerase II and acetylated histone H3K27 binding sites associated with transcriptionally active genes. Cultured mpkCCD cells were treated with either the vasopressin analog dDAVP (0.1 nM) or vehicle for 24 hours, and then processed for chromatin IP. DNA‐protein complexes were immunoprecipitated using either RNA pol II or H3K27ac antibodies. After reversal of crosslinks, DNA fragments were sequenced and mapped to a reference genome. ChIP‐seq data analysis allowed for precise mapping of genomic elements that were significantly enriched or de‐enriched with dDAVP treatment including exonic, intronic, intergenic, and promoter regions. We correlated these changes at the genomic level with corresponding changes in RNA using RNA‐seq analysis of ‘Poly(A)’‐enriched libraries from mpkCCD cells treated with or without dDAVP. Targets enriched in ChIP‐seq and RNA‐seq data sets in the presence of dDAVP included the water channel aquaporin‐2 (Aqp2), genes associated with AQP2 trafficking (Mal2) and AQP2 transcriptional regulation (Elf3), and many others. This study presents a wealth of new data needed for modeling the transcriptional response to vasopressin. Grant Funding Source : Supported by NHLBI